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Research Article

Circ_0026344 restrains metastasis of human colorectal cancer cells via miR-183

, , , , , , & ORCID Icon show all
Pages 4038-4045 | Received 01 Jul 2019, Accepted 26 Aug 2019, Published online: 14 Oct 2019

Figures & data

Figure 1. CCL20 and CXCL8 synergize to facilitate the proliferation and metastasis of cultured CRC cells. SW480 and Caco-2 cells were treated by CCL20 and CXCL8 at concentration of 100 ng/mL for 48 h. Non-treated cells served as control. (A) Cell proliferation measured by SRB assay. (B) Relative migration and (C) invasion tested by transwell chamber. (D) Expression of core proteins in PI3K/AKT/ERK pathway quantified by western blot analysis. (E and F) Semi-quantitative results based on the data from western blot analysis. *p < .05 compared to control group.

Figure 1. CCL20 and CXCL8 synergize to facilitate the proliferation and metastasis of cultured CRC cells. SW480 and Caco-2 cells were treated by CCL20 and CXCL8 at concentration of 100 ng/mL for 48 h. Non-treated cells served as control. (A) Cell proliferation measured by SRB assay. (B) Relative migration and (C) invasion tested by transwell chamber. (D) Expression of core proteins in PI3K/AKT/ERK pathway quantified by western blot analysis. (E and F) Semi-quantitative results based on the data from western blot analysis. *p < .05 compared to control group.

Figure 2. CCL20 and CXCL8 synergize to facilitate the EMT process. SW480 and Caco-2 cells were treated by 100 ng/mL CCL20 and CXCL8 for 48 h. Non-treated cells served as control. (A) Expression of EMT inducers detected by western blot analysis. (B and C) Semi-quantitative results based on the data from western blot analysis. *p < .05 compared to control group.

Figure 2. CCL20 and CXCL8 synergize to facilitate the EMT process. SW480 and Caco-2 cells were treated by 100 ng/mL CCL20 and CXCL8 for 48 h. Non-treated cells served as control. (A) Expression of EMT inducers detected by western blot analysis. (B and C) Semi-quantitative results based on the data from western blot analysis. *p < .05 compared to control group.

Figure 3. CCL20 and CXCL8 synergize to decrease the expression of circ_0026344. (A) SW480 and Caco-2 cells were treated by 100 ng/mL CCL20 and CXCL8 for 48 h. Non-treated cells served as control. Expression of circ_0026344 was measured by qRT-PCR. (B) Expression of circ_0026344 was evaluated by qRT-PCR after the cells were transfected with circ_0026344 overexpression vector (OE-circ) or an empty vector. (C) Expression of circ_0026344 was measured by qRT-PCR after the cells were transfected with circ_0026344 siRNA (si-circ) or the non-targeting negative control (si-NC). Following transfection, relative (D) migration and (E) invasion were estimated by using transwell chamber. *p < .05 compared to the indicated group.

Figure 3. CCL20 and CXCL8 synergize to decrease the expression of circ_0026344. (A) SW480 and Caco-2 cells were treated by 100 ng/mL CCL20 and CXCL8 for 48 h. Non-treated cells served as control. Expression of circ_0026344 was measured by qRT-PCR. (B) Expression of circ_0026344 was evaluated by qRT-PCR after the cells were transfected with circ_0026344 overexpression vector (OE-circ) or an empty vector. (C) Expression of circ_0026344 was measured by qRT-PCR after the cells were transfected with circ_0026344 siRNA (si-circ) or the non-targeting negative control (si-NC). Following transfection, relative (D) migration and (E) invasion were estimated by using transwell chamber. *p < .05 compared to the indicated group.

Figure 4. Silence of circ_0026344 promotes EMT process. (A) SW480 and (C) Caco-2 cells were transfected with circ_0026344 overexpression vector (OE-circ) or circ_0026344 siRNA (si-circ). An empty vector and a non-targeting sequence (si-NC) were transfected as negative controls. Expression of EMT inducers was detected by western blot analysis. (B and D) Semi-quantitative results based on the data from western blot analysis. *p < .05 compared to the indicated group.

Figure 4. Silence of circ_0026344 promotes EMT process. (A) SW480 and (C) Caco-2 cells were transfected with circ_0026344 overexpression vector (OE-circ) or circ_0026344 siRNA (si-circ). An empty vector and a non-targeting sequence (si-NC) were transfected as negative controls. Expression of EMT inducers was detected by western blot analysis. (B and D) Semi-quantitative results based on the data from western blot analysis. *p < .05 compared to the indicated group.

Figure 5. miR-183 is a downstream effector of circ_0026344. (A) SW480 and Caco-2 cells were transfected with circ_0026344 overexpression vector (OE-circ) or circ_0026344 siRNA (si-circ). An empty vector and a non-targeting sequence (si-NC) were transfected as negative controls. Expression of miR-183 was estimated by qRT-PCR. (B) Expression of miR-183 was estimated by qRT-PCR after the cells were transfected with miR-183 mimic or the scrambled negative control (NC). (C) SW480 and (E) Caco-2 cells were transfected with OE-circ alone or in combination with miR-183, after which the expression of core proteins in Wnt/β-catenin pathway was evaluated by western blot analysis. (D and F) Semi-quantitative results based on the data from western blot analysis. *p < .05 compared to the indicated group.

Figure 5. miR-183 is a downstream effector of circ_0026344. (A) SW480 and Caco-2 cells were transfected with circ_0026344 overexpression vector (OE-circ) or circ_0026344 siRNA (si-circ). An empty vector and a non-targeting sequence (si-NC) were transfected as negative controls. Expression of miR-183 was estimated by qRT-PCR. (B) Expression of miR-183 was estimated by qRT-PCR after the cells were transfected with miR-183 mimic or the scrambled negative control (NC). (C) SW480 and (E) Caco-2 cells were transfected with OE-circ alone or in combination with miR-183, after which the expression of core proteins in Wnt/β-catenin pathway was evaluated by western blot analysis. (D and F) Semi-quantitative results based on the data from western blot analysis. *p < .05 compared to the indicated group.